Milbemycin Oxime半抗原设计及其抗体制备

Hapten Design and Antibodies Development of Milbemycin Oxime

通过milbemycin oxime A4、A3(MILO)肟羟基与琥珀酸酐反应,合成了半抗原MILO-5-琥珀酰半酯,采用碳化二亚胺法将其与载体蛋白BSA偶联制备人工抗原.以此人工抗原免疫新西兰大白兔获得多克隆抗体,抗体效价达1:12 800;用此抗体建立的间接竞争酶联免疫吸附(CI-ELISA)检测方法,线性范围在16.00～4 394.18 ng/mL,最低检测限(I10)为6.28 ng/mL.

To produce antigens susceptible to raise antibodies for milbemycin oxime A4 ,A3 （MILO）, the 5-O-succinoylmilbemycin-oxime were bound to bovine serum albumin （BSA） by the carbodiimide activation. Two New Zealand white rabbits were immunized with MILO-BSA and the titers of antiserum had reached 1：12800. A MILO-OVA-based CI-ELISA was conducted with the antiserum, the response range for milbemycin oxime was between 16.00～4394.18ng/mL and the limit of detection was 6.28ng/mL.